Stanley are out of the Checkatrade Trophy after losing 4-0 against Wolves U21s at Molineux.
With all teams - including Crewe and Chesterfield - on three points going into the final round of group games, a win would have guaranteed the Reds a place in the knock-out stages.
However, under their coach Scott Sellars, the Wolves youngsters scored three goals in the first half and added a fourth in the second to progress.
John Coleman made the five allowed changes with Billy Kee returning after a couple of weeks out and Gary Taylor-Fletcher getting the nod to start.
First year professional Ross Sykes started his third successive Checkatrade Trophy game in the heart of the Stanley defence.
Wolves Under 21s opened the scoring when Christian Herc raced onto a through ball from Connor Ronan and his shot hit the inside of the post and bounced into the net on 17 minutes.
Stanley's Sykes had a ‘goal' disallowed for off-side after getting on the end of a Matty Pearson cross while, at the other end, Aaron Chapman pulled off a superb save to stop a second Wolves goal – Bright Enobakhare racing into the area and firing goalwards.
The second did come however on 37 minutes when a corner found Donovan Wilson at the far post and he finished past Chapman.
Wolves made it three from the penalty spot on the stroke of half-time when Sykes was judged to have brought down Morgan Gibbs-White in the area and captain Ronan tucked home the spot kick.
After the re-start, Chapman tipped a Wilson deflected effort around the post as Wolves were in charge.
Coleman put on youth team player Reagan Ogle, 17, for his first team debut on 56 minutes and he slotted into the right back role.
Rommy Boco and Chris Eagles also came on but, on 66 minutes, Enobakhare charged into the area and finished well for number four for Wolves Under 21s.
Jordan Clark forced a save out of Harry Burgoyne with 10 minutes to go but Stanley are out of the competition.
Stanley: Chapman, Pearson, Hughes, Sykes, Lacey; McConville, Conneely, Hewitt, Clark, Taylor-Fletcher, Kee
Stanley subs: Parish, Davies, Beckles, Gornell, Boco, Ogle, Eagles